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Objective. This study aims to evaluate the efficacy of various conventional synthetic DMARDs, including Tripterygium wilfordii Hook F (TwHF) for treating rheumatoid arthritis (RA) by network meta-analysis. Methods. We retrieved the related literature from online databases and supplemented it by using a manual retrieval method. Data was extracted from the literature and analyzed with STATA software. Results. A total of 21 trials (5,039 participants) were identified. Assessment of ACR20 response found that TwHF combined with methotrexate (MTX) had the greatest probability for being the best treatment option among the treatments involved, while TwHF used singly was second only to TwHF combined with MTX. Assessment of ACR50 response found that TwHF combined with MTX ranked second in all treatment options after cyclosporine A (CsA) combined with leflunomide (LEF) and TwHF alone, followed by TwHF combined with MTX. Assessment of ACR70 response found that CsA combined with LEF ranked first, TwHF combined with LEF ranked second, TwHF combined with MTX ranked third, and TwHF used singly ranked fourth. In the safety analysis, TwHF had the least probability of adverse event occurrence, followed by TwHF combined with MTX, which ranked first and second, respectively. Conclusion. Compared with the current csDMARDs for treating RA, the efficacy of TwHF was clear, and TwHF combined with MTX performed well under various endpoints. In the future, large, rigorous, and high-quality RCTs are still needed to confirm the benefits of TwHF therapy on RA.

This study aims to evaluate the efficacy of various conventional synthetic DMARDs, includingWe retrieved the related literature from online databases and supplemented it by using a manual retrieval method. Data was extracted from the literature and analyzed with STATA software.A total of 21 trials (5,039 participants) were identified. Assessment of ACR20 response found that TwHF combined with methotrexate (MTX) had the greatest probability for being the best treatment option among the treatments involved, while TwHF used singly was second only to TwHF combined with MTX. Assessment of ACR50 response found that TwHF combined with MTX ranked second in all treatment options after cyclosporine A (CsA) combined with leflunomide (LEF) and TwHF alone, followed by TwHF combined with MTX. Assessment of ACR70 response found that CsA combined with LEF ranked first, TwHF combined with LEF ranked second, TwHF combined with MTX ranked third, and TwHF used singly ranked fourth. In the safety analysis, TwHF had the least probability of adverse event occurrence, followed by TwHF combined with MTX, which ranked first and second, respectively.Compared with the current csDMARDs for treating RA, the efficacy of TwHF was clear, and TwHF combined with MTX performed well under various endpoints. In the future, large, rigorous, and high-quality RCTs are still needed to confirm the benefits of TwHF therapy on RA.

Health effects resulting from the smoke of carbonyl compounds (aldehydes and ketones) and metal-containing incense particles at temples during incense burning periods were evaluated at temple A (without incense reduction activities) and B (with incense reduction activities), Nantou County, in 2018. The predominant size fractions of particles were PM1, PM1–2.5, and PM2.5–10 at both temples. The total particle mass at temple A was approximately 1.1 times that of temple B due to incense reduction at temple B. The most abundant metal elements in all particle size fractions at both temples were Fe, Al, and Zn. Metal species of incense smoke are divided into three groups by hierarchical cluster analysis and heatmaps, showing higher metal contents in groups PM1, PM18–10, and PM18–2.5 at temple A. In contrast, higher metal contents were observed in PM18–10 and PM2.5–1 at temple B. Most of the carbonyl species were formaldehyde and acetaldehyde, released during incense burning periods, with concentrations ranging from 6.20 to 13.05 μg/m3 at both temples. The total deposited fluxes of particle-bound metals at temples A and B were determined to be 83.00% and 84.82% using the International Commission on Radiological Protection (ICRP) model. Health-risk assessments revealed that the risk values of metals and carbonyls were above recommended guidelines (10−6) at temple A. Since worshippers and staff are exposed to incense burning environments with poor ventilation over a long period, these toxic organic compounds and metals increase health risks in the respiratory tract. Therefore, incense reduction is important to achieve healthy temple environments.

MAP30 (Momordica antiviral protein 30kD) is a single-chain Ⅰ-type ribosome inactivating protein with a variety of biological activities, including anti-tumor ability. It was reported that MAP30 would serve as a novel and relatively safe agent for prophylaxis and treatment of liver cancer. To determine whether adding two tumor targeting peptides could improve the antitumor activities of MAP30, we genetically modified MAP30 with an RGD motif and a EGFRi motif, which is a ligand with high affinity for αvβ3 integrins and with high affinity for EGFR. The recombinant protein ELRL-MAP30 (rELRL-MAP30) containing a GST-tag was expressed in E. coli. The rELRL-MAP30 was highly expressed in the soluble fraction after induction with 0.15 mM IPTG for 20 h at 16 °C. The purified rELRL-MAP30 appeared as a band on SDS–PAGE. It was identified by western blotting. Cytotoxicity of recombinant protein to HepG2, MDA-MB-231, HUVEC and MCF-7 cells was detected by MTT analysis. Half maximal inhibitory concentration (IC50) values were 54.64 μg/mL, 70.13 μg/mL, 146 μg/mL, 466.4 μg/mL, respectively. Proliferation inhibition assays indicated that rELRL-MAP30 could inhibit the growth of Human liver cancer cell HepG2 effectively. We found that rELRL-MAP30 significantly induced apoptosis in liver cancer cells, as evidenced by nuclear staining of DAPI. In addition, rELRL-MAP30 induced apoptosis in human liver cancer HepG2 cells by up-regulation of Bax as well as down-regulation of Bcl-2. Migration of cell line were markedly inhibited by rELRL-MAP30 in a dose-dependent manner compared to the recombinant MAP30 (rMAP30). In summary, the fusion protein displaying extremely potent cytotoxicity might be highly effective for tumor therapy.

Mesenchymal stem cells have been widely studied to promote local bone regeneration of osteonecrosis of the femoral head (ONFH). Previous studies observed that dimethyloxaloylglycine (DMOG) enhanced the angiogenic and osteogenic activity of mesenchymal stem cells by activating the expression of hypoxia inducible factor-1α (HIF-1α), thereby improving the bone repair capacity of mesenchymal stem cells. In the present study, it was investigated whether DMOG could increase the bone repair capacity of adipose-derived stem cells (ASCs) in the treatment of ONFH. Western blot analysis was performed to detect HIF-1α protein expression in ASCs treated with different concentrations of DMOG. The results showed DMOG enhanced HIF-1α expression in ASCs in a dose-dependent manner at least for 7 days. Furthermore, DMOG-treated ASCs were transplanted into the necrotic area of a rabbit model of ONFH to treat the disease. Four weeks later, micro-computed tomography (CT) quantitative analysis showed that 58.8±7.4% of the necrotic area was regenerated in the DMOG-treated ASCs transplantation group, 45.5±3.4% in normal ASCs transplantation group, 25.2±2.8% in only core decompression group and 10.6±2.6% in the untreated group. Histological analysis showed that transplantation of DMOG-treated ASCs clearly improved the bone regeneration of the necrotic area compared with the other three groups. Micro-CT and immunohistochemical analysis demonstrated the revasculation of the necrotic area were also increased significantly in the DMOG-treated ASC group compared with the control groups. Thus, it is hypothesized that DMOG could increase the bone repair capacity of ASCs through enhancing HIF-1α expression in the treatment of ONFH.

Chitosan (CTS) and mesoporous calcium silicate (MCS) have been developed for bone defect healing; however, their bone regeneration capacity still does not satisfy the patients with bone diseases. Gadolinium (Gd) is accumulated in human bones, and plays a beneficial role in regulating cell performance and bone regeneration. We firstly constructed Gd-doped MCS/CTS (Gd-MCS/CTS) scaffolds by a lyophilization technology. The interconnected arrangement of CTS films lead to forming macropores by using ice crystals as templates during the lyophilization procedure, and the Gd-MCS nanoparticles dispersed uniformly on the macropore walls. The biocompatible chemical components and hierarchical pores facilitated the attachment and spreading of rat bone marrow-derived mesenchymal stem cells (rBMSCs). Interestingly, the Gd dopants in the scaffolds effectively activated the Wnt/β-catenin signaling pathway, resulting in excellent cell proliferation and osteogenic differentiation capacities. The osteogenic-related genes such as alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2) and collagen type1 (COL-1) were remarkably up-regulated by Gd-MCS scaffolds as compared with MCS scaffolds, and their expression levels increased in a positive correlation with Gd doping amounts. Moreover, in vivo rat cranial defect tests further confirmed that Gd-MCS/CTS scaffolds significantly stimulated collagen deposition and new bone formation. The exciting finding suggested the beneficial effects of Gd3+ ions on osteogenic differentiation and new bone regeneration, and Gd-MCS/CTS scaffolds can be employed as a novel platform for bone defect healing.

Detailed mechanisms of W hi B - l ike (Wbl) proteins involved in antibiotic biosynthesis and morphological differentiation are poorly understood. Here, we characterize the role of WblA ch , a Streptomyces chattanoogensis L10 protein belonging to this superfamily. Based on DNA microarray data and verified by real-time quantitative PCR (qRT-PCR), the expression of wblA ch was shown to be positively regulated by AdpA ch . Gel retardation assays and DNase I footprinting experiments showed that AdpA ch has specific DNA-binding activity for the promoter region of wblA ch . Gene disruption and genetic complementation revealed that WblA ch acts in a positive manner to regulate natamycin production. When wblA ch was overexpressed in the wild-type strain, the natamycin yield was increased by ∼30%. This provides a strategy to generate improved strains for natamycin production. Moreover, transcriptional analysis showed that the expression levels of whi genes (including whiA , whiB , whiH , and whiI ) were severely depressed in the Δ wblA ch mutant, suggesting that WblA ch plays a part in morphological differentiation by influencing the expression of the whi genes.

To explore the potential association between the serum levels of 25-hydroxyvitamin D [25(OH)D] and carotid atherosclerosis in patients with type 2 diabetes.Three hundred and fifty patients with type 2 diabetes were enrolled in this study in Shanghai, China. B-mode ultrasound was used to detect carotid plaques as indicators of atherosclerosis and measure carotid artery intima-media wall thickness (C-IMT) at two sites of carotid artery. Subjects were divided into group A (patients with carotid plaques) and group B (patients without carotid plaques) and be assessed clinically. Serum levels of 25(OH)D and other clinical parameters were measured. Multivariate logistic regression was performed to find predictors of carotid atherosclerosis in the entire group.The levels of serum 25(OH)D were lower in group A than in group B[19.60 (13.30-25.73) vs 23.19 (18.10-30.06)ng/ml, P0.001]. The C-IMT levels [(1.00±0.17 vs 0.88±0.20)mm, Ptrend0.001] and proportion of people with carotid plaques(44/88 vs 20/87, Ptrend0.001) in the lowest quartile of 25(OH)D were higher than in the highest quartile. Vitamin D concentrations were inversely associated with HbA1c in women(r=-0.194, P=0.006), and C-IMT in men(r=-0.409, P0.001). Logistic regression analysis showed age, male sex, current smoke, history of hypertension, SBP, LDL-C and lg[25(OH)D] (OR: 0.924, 95%CI: 0.893-0.955, P0.001) were independently associated with the presence of carotid plaques in T2DM.Serum vitamin D level is significantly and independently associated with carotid atherosclerosis in patients with T2DM in Shanghai, China.

Salinomycin, a polyether antibiotic produced by Streptomyces albus, is widely used in animal husbandry as an anticoccidial drug and growth promoter. Situated within the salinomycin biosynthetic gene cluster, slnR encodes a LAL-family transcriptional regulator. The role of slnR in salinomycin production in S. albus was investigated by gene deletion, complementation, and overexpression. Gene replacement of slnR from S. albus chromosome results in almost loss of salinomycin production. Complementation of slnR restored salinomycin production, suggesting that SlnR is a positive regulator of salinomycin biosynthesis. Overexpression of slnR in S. albus led to about 25 % increase in salinomycin production compared to wild type. Quantitative RT-PCR analysis revealed that the expression of most sal structural genes was downregulated in the ΔslnR mutant but upregulated in the slnR overexpression strain. Electrophoretic mobility gel shift assays (EMSAs) also revealed that SlnRDBD binds directly to the three intergenic regions of slnQ-slnA1, slnF-slnT1, and slnC-slnB3. The SlnR binding sites within the three intergenic regions were determined by footprinting analysis and identified a consensus-directed repeat sequence 5′-ACCCCT-3′. These results indicated that SlnR modulated salinomycin biosynthesis as an enhancer via interaction with the promoters of slnA1, slnQ, slnF, slnT1, slnC, and slnB3 and activates the transcription of most of the genes belonging to the salinomycin gene cluster but not its own transcription.

Simultaneous bilateral fractures of the femoral neck are rare injuries, which are reportedly induced by low-speed energy with predisposing factors including systemic diseases, medications and eclamptic seizures. Those caused by high energy are even rarer. High energy-induced bilateral fractures of the femoral neck conceive of high incidence of mortality and present great challenges in the early management. We report one case of a 52-year-old man with simultaneous bilateral fractures of the femoral neck which resulted from a motor pedestrian accident. One-stage closed reduction and internal fixation was done following the emergent resuscitation and neurosurgical management for concomitant brain injuries. The fractures united. There was no pain in the hips, and they had a normal range of motion. The treatment protocol, mechanism of the injury and possible postoperative complications were discussed to expand a comprehensive understanding about these infrequent types of fractures.

The aim of this study was to evaluate the risk factors of mild cognitive impairment (MCI) in middle-aged patients with type 2 diabetes (T2DM).Montreal Cognitive Assessment (MoCA) was applied as cognition assessment implement. One hundred and fifty-seven middle-aged type 2 diabetic patients were enrolled in this cross-section study (age 40~69, mean age 55 ± 7). There were 93 patients with MCI (MoCA score26) in MCI group and 64 with normal cognitive function (MoCA score ≥ 26) in control group. Information of history of disease, family history, data of BMI, WHR, HbA1c, FINS, C-Peptide (C-P), SBP, DBP, blood lipid (TG, TC, LDL-C, HDL-C and carotid ultrasound (carotid IMT, carotid resistance index [RI]) was collected.There were significant differences in the rate of patients with hypertension ([40.63 vs. 58.06%], P=0.026), duration of diabetes mellitus ([3.09 ± 4.04 y vs. 4.80 ± 4.94 y], P=0.024), C-P ([2.79 ± 1.09 ng/ml vs. 2.26 ± 1.00 ng/ml], P=0.008), Max C-IMT ([0.81 ± 0.15 mm vs. 0.91 ± 0.15 mm], P0.001), Min C-RI (0.71 ± 0.06 vs. 0.68 ± 0.06, P0.05), and no significant differences in the duration of hypertension and hyperlipidemia, BMI, WHR, HbA1c, SBP, DBP and blood lipid between control group and MCI group. MoCA scores were positively correlated with C-P (r=0.252, P=0.005), and negatively correlated with the history of hypertension (r=-0.244, P=0.002), duration of DM (r=-0.161, P=0.044), Max C-IMT (r=-0.253, P=0.005) and Min C-RI (r=-0.183, P=0.023). Multiple regression analysis showed that history of hypertension (Beta=-0.267, P=0.002), C-P (Beta=0.281, P=0.001) and Min C-RI (Beta=-0.221, P=0.011) were significantly independent determinants for the MoCA scores.The longer duration of diabetes, history of hypertension, lower serum C-P levels, thickened C-IMT and higher C-RI could be risk factors of MCI in type 2 diabetic patients. This finding could have an important impact on the management of cognitive decline in diabetic patients.

Ethnopharmacological relevanceAstragaloside IV and tetramethylpyrazine have been extensively used in the cardio-cerbrovascular diseases of medicine as a chief ingredient of glycoside or alkaloid formulations for the treatment of stroke and myocardial ischemia diseases.Aim of the studyTo investigate the effects of astragaloside IV (ASG IV) and tetramethylpyrazine (TMPZ) on cerebral ischemia-reperfusion (IR) injury model in rat model.Materials and methodsRats were randomly divided into the following five groups: sham group, IR group and treatment group including ASG IV, ASG IV–TMPZ and nimodipine treatment. The therapeutic effect was evaluated by micro-positron emission tomography (Micro-PET) using 18F-fluoro-2-deoxy-d-glucose. The neurological examination, infarct volume and the levels of oxidative stress- and cell apoptosis-related molecules were assessed.ResultsMicro-PET imaging showed that glucose metabolism in the right hippocampus was significantly decreased in the IR group compared to the sham group (P

Summary Background It is controversial whether an early reduction and internal fixation can reduce the occurrence of femoral neck fracture-induced osteonecrosis of the femoral head (ONFH). This prospective study was designed to reflect the relationship between injury-to-surgery interval (ISI) and traumatic ONFH based on a canine model of femoral neck fractures. Material/Methods Twenty-four dogs were equally divided randomly into 3 groups. A lateral L-shape approach centered left great trochanter was used for exposure of the femoral neck. A low-speed drill was used for making displaced fractures in the narrow femoral neck, with the femoral head kept in situ with ligamentum teres intact. In Group A, the fracture was immediately reduced and fixed with 3 parallel pins; while the operation was done 3 days later in Group B, and 3 weeks later in Group C. Another 2 dogs had their fractures untreated. Postoperatively, all dogs were fed separately and received regular x-ray examination. Left femoral heads were harvested for histological examination with a postoperative follow-up of 3.5 months. Results The canine model of femoral neck fractures could be achieved successfully. Radiological signs of post-fracture ONFH could not be detected at intervals of 2 weeks, 4 weeks, 1 month and 2 months. Histologically, there were 2 cases with ONFH in Group A, 1 case in Group B, and 2 cases in Group C. The difference had no statistical significance. For untreated fractures, obvious ONFH could be found radiologically. Conclusions A shorter ISI may not reduce the incidence of fracture-induced ONFH, which suggests that intrinsic factors play an important role in the occurrence of ONFH.

The correlation of PK/PD for Danhong Injection (DHI) was investigated by the least square method in a rat model of cerebral ischemia/reperfusion. [Methods] Sprague-Dawley rats were randomly divided into 4 equal groups (each n = 6) (sham group), cerebral IR damage alone (model group), IR model treated with DHI (treatment group), and IR model treated with nimodipine (positive group). The drugs concentration of Danshensu (DSS) and Hydroxysafflor yellow A (HSA) in Plasma were measured by HPLC at 5, 15, 30, 60, 120, 180, 240, and 360 min. The levels of IL-1s in Plasma were measured by method of enzyme-linked immunosorbent assay. Curves were fit using non-linear least squares methods. The results showed that levels of IL-1s were decreased after 60, 120, 180, 240, and 360 min (P

This article previously published in Volume 15 Issue 10 of this journal in October 2016 has been retracted in line with the guidelines from the Committee on Publication Ethics (COPE, http://publicationethics.org/resources/guidelines) Retracted: Ma Z, Yin W, Hu G, Zhu Z, Huang Z. Effect of Paris polyphylla extract on second-degree burns in rats. Trop J Pharm Res 2016; 15(10):2131-2135 doi: http://dx.doi.org/10.4314/tjpr.v15i10.11 From the Editor Our attention was drawn to the falsification of the data published in this article which was confirmed. The corresponding author, Zhi-jian Huang, failed to respond to communication in this respect. 26 January 2018

ObjectiveTo investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta (IL-1β) and neuropeptide Y (NPY) based on pharmacodynamics in rats.MethodsThe animal model was built by transient middle cerebral artery occlusion (MCAO). The method for evaluating the concentrations of the FA-Pr-Al components in rat plasma was established by using HPLC and the expression levels of IL-1β and NPY were determined by ELISA. A new mathematics method of the trend of percentage rate of change (PRC) was used to assess the correlation between pharmacokinetics (PK) and pharmacodynamics (PD).ResultsFA-Pr-Al in combination reduced neurological deficits, decreased infarct volume and inhibited the expression levels of IL-1β and NPY (all P

Based on the feature of Fermentation Engineering Course, Study on student-centered classroom teaching by new teaching modes and methods to stimulate students’ learning motivation and develop students’ innovating ability. Combines problem based learning (PBL) with case based learning (CBS) teaching methods. In the process of teaching, guiding the students to think actively through questions, and participation in class discussion, courseware, simulation teaching, etc. and guiding students to analyze and solve problems, construction guide classroom teaching model. The resultsshow that the student’s initiative, enthusiasm and learning effectwereimproved obviously. And the way of teaching achieved good teaching results, which not only highlightedthe student-centered teaching philosophy, but also learnedthe advantages of a wide range of teaching methods.

An improved procedure for the fermentation and purification of human epidermal growth factor (hEGF) was developed. Recombinant Escherichia coli HB-101 [lacUV5omp08hEGF] harboring plasmid lacUV5omp08hEGF encoding hEGF was used in fermentation to increase levels of hEGF. Medium composition, and the levels of inoculum, inducer (isopropyl-beta-D-thiogalactoside) and ampicillin were optimized with respect to volumetric fermentation of hEGF. As a result, the hEGF concentration reached a high value of 242 mg l(-1) and the amount of heterogeneous protein decreased by 62% compared with that before optimization in batch fermentation. High-quality hEGF was purified from the fermentation culture by centrifugation, salting-out, resuspension, recentrifugation and finally gel chromatography on a Grad-iFrac System using Sephadex G-50 superfine. The purity of hEGF and the total yield were more than 94% and higher than 36%, respectively, and SDS-PAGE of the purified hEGF demonstrated a single band corresponding to an hEGF standard. In particular, a very important phenomenon was found, i.e. that the amount of heterogenous protein in fermentation broths cultured in media with high concentrations of lactose is far less than that cultured in media with high concentrations of glucose.

Purpose: To investigate the healing effect of Paris polyphylla extract (PPE) on second-degree burns in rats. Methods : Male Sprague Dawley (SD) rats, weighing 200 – 220 g, were subjected to deep seconddegree skin burns by electrical scald instrument. The animals were divided into three groups as follows: (1) second-degree burn model (control) group, (2) burn model treated with 1 % silver sulfadiazine (SSD) group, and (3) burn model treated with 120 mg·mL -1 PPE group. On days 3, 7 and 14 following the administration of the drug/extract, wound area and histopathological changes in rat epidermis were evaluated for the three groups. The minimum inhibitory concentration (MIC) of PPE on Staphyloccocus aureus , Pseudomonas aeruginosa and Escherichia coli were also assessed. Results: On day 14, the mean wound area of PPE treatment group (0.21 ± 0.04 cm 2 ) was significantly smaller than that of the control rats (2.78 ± 0.18 cm 2 , p < 0.01). Histological results indicate that inflammatory cells disappeared and were replaced by new granulation tissue in the group treated with 120 mg·mL -1 PPE by day 14. Compared with SSD group rats, the inflammatory cells and fibroblast and granulation tissues of burnt rats with burns and treated with 120 mg·mL -1 PPE decreased significantly. The antibacterial data revealed that the MIC of PPE against S. aureus , P. aeruginosa and E. coli was 2.35, 8.2 and 4.70 mg·mL -1 , respectively. Conclusion: Paris polyphylla is an effective medicinal herb that holds promise for the treatment of second-degree burns. Keywords: Paris polyphylla, Second-degree burns, Wound healing, Antibacterial, Inflammatory cells, Granulation tissues, Silver sulfadiazine

To explore the antimicrobial resistance of nosocomial Gram-negative bacilli across China.A total of 1247 consecutive and non-repetitive Gram-negative bacilli were isolated from 13 Chinese teaching hospitals from March to August 2012. All isolates were sent to a central laboratory for reidentification and susceptibility testing. The minimal inhibitory concentration (MICs) of meropenem and other antibacterial agents were determined by agar dilution method. And the data were analyzed with WHONET-5.6 software.The activity of antimicrobial agents against Enterobacteriaceae was in the following descending order of susceptibility rate: meropenem (97.5%, 849/871) , amikacin (94.5%, 823/871) , imipenem (93.6%, 815/871) , ertapenem (92.9%, 809/871) , piperacillin/tazobactam (89.9%, 783/871) , cefoperazone/sulbactam (83.5%, 727/871) , cefepime (78.1%, 680/871) , polymyxin B (77.0%, 670/871) , cefiazidime (69.6%, 606/871) , levofloxacin (69.2%, 603/871) , ciprofloxacin (63.6%, 554/871) , minocyline (63.1%, 550/871) , ceftriaxone (55.7%, 485/871) , cefotaxime (54.2%, 472/871) and cefoxitin (51.4%, 448/871) . The prevalence of extended-spectrum beta-lactamase (ESBL) was 64.3% (117/182) in Escherichia coli (E. coli) and 32.1% (60/187) in Klebsiella pneumonia (K. pneumoniae) . The sensitivities of E. coli to meropenem and imipenem were 100%. And over 90% of E. coli was sensitive to ertapenem, amikacin, piperacillin/tazobactam and polymyxin B. However, over 60% of E. coli was resistant to ciprofloxacin, levofloxacin, ceftriaxone and cefotaxime. The susceptibility of K. pneumoniae to meropenem, imipenem, amikacin and polymyxin B maintained at over 90%. The activities of antimicrobial agents against E. cloacae, E. aerogenes and Citrobacter freundii were in the following descending order of susceptibility rate: meropenem (96.0%-100%) , imipenem (96.0%-100%) , polymyxin B (95.8%-100%) , amikacin (92.2%-100%) , ertapenem (85.6%-93.3%) , cefepime (77.8%-93.3%) , cefoperazone/sulbactam (78.4%-90.0%) and piperacillin/tazobactam (65.0%-89.8%) . The most susceptible agent against Acinetobacter baumannii (A. baumannii) was polymyxin B (100%) . The susceptibilities of A.baumannii to imipenem, meropenem and minocyline were 37.8% (65/172) , 36.0% (62/172) and 62.8% (108/172) respectively. The most active agents against Pseudomonas aeruginosa (P. aeruginosa) were polymyxin B (97.2%, 173/178) , followed by amikacin (89.3%, 159/178) and cefiazidime (83.7%, 149/178) . Clinical and Laboratory Standards Institute revised P.aeruginosa susceptibility standard in 2012. The sensitivity of piperacillin/tazobactam changed from 83.7% (149/178) to 77.5% (138/178) . The sensitivity of meropenem decreased from 78.1% ( 139/178 ) to 71.3% ( 127/178 ) while that of imipenem declined from 69.7% (124/178) to 59.6% (106/178) . The prevalence of multi-drug resistant A. baumannii and P. aeruginosa were 65.7% (113/172) and 9.0% (16/178) respectively.Carbapenems remain highly active against Enterobacteriaceae. Increasing resistance of A. baumannii to all antimicrobial agents is noted. New breakpoint to P.aeruginosa has obvious effects on antimicrobial sensitivity.

Hypovitaminosis D is highly prevalent in type 2 diabetes. The aim of this study is to determine the serum levels of 25-hydroxyvitamin D [25(OH)D] in type 2 diabetic patients with and without mild cognitive impairment (MCI), and examine the relationship of 25(OH)D and MCI with other clinical factors. One hundred and sixty-five diabetic patients were enrolled in this study. Among whom, 95 patients were considered as MCI [Montreal Cognitive Assessment score (MoCA) < 26] and the other 70 as no MCI (MoCA ≥ 26). Subjects were assessed clinically. Diabetic patients with MCI had a longer duration of DM, fewer years of education, elevated fasting blood glucose (FBG), resistant index (RI) of carotid, and lower levels of 25(OH)D {[17.35 (13.02-25.92) vs 28.00 (19.67-34.30)] ng/ml, P < 0.001}. The MoCA score was positively correlated with log10[25(OH)D], education year, and inversely correlated with duration of DM, history of hypertension, intima-media thickness (IMT), FBG, max-RI, and min-RI. Log10[25(OH)D] was positively correlated with MoCA score, and inversely correlated with IMT, in multivariate regression analysis adjusted for age, sex, and education year, 25(OH)D (β = 0.210, P = 0.003), history of hypertension (β = -0.191, P = 0.007), IMT (β = -0.194, P = 0.007), and FBG (β = -0.157, P = 0.026) independently predicted MoCA score. In conclusion, our results suggest that levels of serum 25(OH)D are inversely associated with the cognitive impairment in diabetic patients. Vitamin D may be a potential protective factor for cognitive impairment in patients with type 2 diabetes.

To observe the effect and mechanism of Yiqi Tongluo Jiedu capsule aganist cerebral ischemia reperfusion injury, the SD rats were randomly divided into following groups: sham-operated group, model group, the group of low, medium and high dose of Yiqi Tongluo Jiedu capsule, and nimodipine group. Using focal middle cerebral artery embolization (MCAO) model, following items were observed: symptoms of neurological deficit score; infarct volume; activity of SOD, content of MDA and NO, activity of NOS of ischemic brain tissue; Bcl-2 and Bax protein expression; content of IL-1beta, IL-6 and TNFalpha in serum; IL-1beta mRNA expression of ischemic brain tissue. Results showed that Yiqi Tongluo Jiedu capsule could significantly reduce the symptoms of neurological deficits, promote the recovery symptoms of neurological deficits; narrow infarct volume of brain tissue obviously, reduce the percentage of infarct volume; raise activity of SOD, reduce content of MDA and NO, reduce activity of NOS; increase Bcl-2 protein, reduce Bax expression; reduce content of IL-1beta, IL-6 and TNFa in serum; reduce IL-1beta mRNA expression of ischemic brain tissue. Yiqi Tongluo Jiedu capsule has significant protective effects against ischemic brain injury, it has significant anti-apoptotic, antioxidant and anti-inflammatory effects.

Fritillaria thunbergii Miq., known as the bulbous plants of the genus fritillaria, produces a large amount of sterols. Homology based polymerase chain reactions (PCRs) with degenerate primers designed from the conserved sequences among the known cycloartenol synthase (CAS) resulted in cloning of a CAS from the young leaves of F. thunbergii Miq. A putative cycloartenol synthase cDNA (FtCAS) consists of a 2271 bp open reading frame, which encodes for 756 amino acids. The deduced amino acid sequence shows 82% homology to cycloartenol synthases from Dioscorea zingiberensis. Heterologous expression of the FtCAS in the methylotrophic yeast, Pichia pastoris, resulted in production of cycloartenol. Our results indicate that FtCAS encode cycloartenol synthase.Key words: Fritillaria thunbergii Miq., cycloartenol synthase, cDNA cloning, 2,3-oxidosqualene cyclase.

Correct genotype identification of medicinal plant material remains important for Chinese traditional medicine. Limitations of chemical and morphological approaches for authentication have generated need for newer methods in discrimination of famous-region herbal medicines. The present study was carried out to develop DNA based markers for identification of Chinese famous-region drug Ophiopogon japonicus (L.f.) Ker-Gawl. Two specific RAPD fragments R01 (268bp) and R02 (1364bp) for famous-region drug Ophiopogon japonicus (L.f.) Ker-Gawl were identified by Random Amplified Polymorphic DNA (RAPD). Sequence Characterized Amplified Region (SCAR) markers (248bp & 631bp) were developed from the RAPD amplicon. The SCAR markers were found useful for identification of genuine regional Ophiopogon japonicus ( L.f. ) Ker-Gawl in four different populations.

Osteonecrosis of the femoral head (ONFH) is a common and severe complication following femoral neck fractures in young adults. Despite significant advances in surgical techniques, radiological evaluation and comprehensive treatment for the prevention of ONFH, the incidence of traumatic ONFH has remained unchanged at approximately 20% in recent decades. The injury-to-surgery interval is considered as a principal factor affecting the occurrence of ONFH, and traditionally, femoral neck fractures are treated emergently. However, the relationship between the injury-to-surgery interval and ONFH occurrence is poorly understood, and previous reviews have not provided a precise explanation due to the lack of strict selection criteria for studies.We reviewed previously published articles and included in current systematic review those studies with accurate multivariate analyses that included age, fracture type, operation method, follow-up, ONFH occurrence and injury-to-surgery interval.Six case studies were included and reevaluated. The studies included 263 hips for final analysis, with an overall incidence of postfracture ONFH of 17.5%. Patients were categorized into groups of less/more than 8 h, less/more than 24 h, less/more than 48 h and less/more than 3 weeks based on the individual injury-to-surgery interval. The postfracture ONFH incidence ranged from 13.3% (8 weeks) to 21.7% (3 weeks). Operations performed within 3 weeks of injury resulted in a lower ONFH incidence compared with operations performed after 3 weeks; however, this difference was not statistically significant. The ONFH incidence remained relatively stable when the operations were performed within 3 weeks of injury.The injury-to-surgery interval did not significantly affect the incidence of postoperative ONFH.

Pain is the core and basic problem in the treatment of osteonecrosis of the femoral head (ONFH). However, it is unclear about the status of pain relief following ONFH treated by free vascularized fibular grafting (FVFG) and the level of pain relief contributed to clinical results. Therefore, we designed a consecutive and prospective study to investigate post-operative pain relief in the treatment of osteonecrosis of the femoral head by FVFG.One hundred and fifty-one patients with unilateral osteonecrosis of the femoral head were enrolled consecutively for current prospective study from January to August of 2006. Patients were managed by modified technique of free vascularized fibular grafting. Pre-operative, post-operative Harris hip score (HHS) and Harris pain score (HPS) were recorded and compared statistically, meanwhile, correlation between disease severity and Harris hip score, Harris pain score were revealed.All patients had an average follow-up of 54.3 months. Post-operative Harris hip score could be improved from 73.7 to 83.5 averagely in stage-II patients, 64.6 to 78.9 in stage-III, and 53.6 to 72.4 in stage-IV. As for Harris pain score, it was elevated from 28.8 to 38.6 in stage-II patients, 25.5 to 36.6 in stage-III, and 21.8 to 34.2 in stage-IV. Taken together, HHS was improved from 67.7 to 80.3 (ΔHHS = 12.6), and HPS was improved from 26.6 to 37.3 (ΔHPS = 10.7) averagely.Harris pain score could be employed to monitor prognosis of osteonecrosis of the femoral head treated by free vascularized fibular grafting. Improvement of HPS was the heaviest contributor to elevation of HHS, and both of them conceived of a close relationship with disease severity.

The purpose of this study was to establish an absolute quantitative method to detect IL-1β and Caspase-3 gene expressions in rat brain after cerebral ischemia-reperfusion (I/R) using real-time PCR. Rats were randomized into the following groups: sham operation group, model group (cerebral I/R group), astragaloside IV (AST IV) group, chuanxiongzine-AST IV group and nimodipine group (n = 10 in each group). The rats in all the groups except sham operation group were subjected to cerebral I/R treatment. Sham operation and model groups were treated by normal saline (5 mL/kg). AST IV, chuanxiongzine-AST IV and nimodipine groups received 20 mg/kg AST IV, 10 mg/kg chuanxiongzine plus 20 mg/kg AST IV, and 10 mg/kg nimodipine treatments, respectively. The administrations of drugs were performed with intraperitoneal injections at 0 and 12 h, 1 d, 2 d, 3 d, till to 7 d after I/R. A real-time quantitative PCR assay was developed for absolute quantification of the expressions of IL-1β and Caspase-3 genes. The absolute quantification approach relies on the construction of an accurate standard curve. Thus, two plasmids which contained rat IL-1β and Caspase-3 genes respectively were constructed. The cloned circular plasmids were then quantified using a spectrophotometer and used as standards. Standard curves were generated, and the copy numbers of IL-1β and Caspase-3 mRNA isolated from I/R-damaged brain tissue were also calculated by SYBR Green I dye method using specific primers. The results showed that melting curves exhibited sharp peaks, and PCR product also generated prominent band with expected size in agarose gel electrophoresis, which validated the optimization of the selected primer sets of IL-1β and Caspase-3 genes. The optimal annealing temperatures of IL-1β and Caspase-3 genes were 59 °C and 61.2 °C, respectively. Real-time PCR results showed that the expression of IL-1β and Caspase-3 genes in the model group was significantly elevated compared to that in the sham operation group. However, compared to those in the model group, IL-1β and Caspase-3 gene expressions were obviously decreased in AST IV, chuanxiongzine-AST IV and nimodipine groups. Especially in chuanxiongzine-AST IV group, those two genes showed the most significant expression down-regulation. These results suggest the absolute quantitative method established in the present study is capable of detecting the changes of IL-1β and Caspase-3 gene expressions in rat brain damaged by I/R.

Summary Background The lack of an experimental animal model that can reliably mimic all stages of osteonecrosis of the femoral head has hindered progress toward the successful prevention and treatment of the disease. Material/Methods A goat model of osteonecrosis of the femoral head (ONFH) was established and observed from the early to the intermediate-to-late stage of mechanical failure. Absolute alcohol was injected slowly into the center of bilateral femoral heads in 12 adult Small Tail Han goats. Postoperatively, the femoral heads were harvested and examined using macrostructural and histological analyses and radiographic and MRI examinations at weeks 4, 8, 12, and 25. Results Macrostructural and radiographic examinations revealed that the contour of both femoral heads was deformed slightly at 12 weeks, but a contour deformation with joint space narrowing was observed at 25 weeks. Histologically, a strong concordance with the natural history of ONFH in humans was found. The present model demonstrated bone trabeculae, marrow necrosis, a reconstruction deficiency and destruction of the microcirculation. Conclusions Among quadrupedal models, the goat model of ONFH, which is induced by a single injection of absolute alcohol, may be suitable and valuable for the evaluation of various therapeutics and side effects in the treatment of ONFH.

Outcomes concerning treatment of severely open fractures of the tibia have achieved great success with the advent of powerful antibiotics, superb microsurgery techniques, and advanced stabilization instruments. However, severe pollution in the wound and high comminution of the fragments may restrict application of internal or external fixators. The condition could appear more complicated if osteoporosis and polytrauma are present or if those special instruments are unavailable.We tried using a surgical suture to stabilize severely open fractures of the tibia in the elderly with osteoporosis. A 65-year-old woman with a severely open fracture of the tibia was treated with a surgical suture, and the result was satisfactory in early period. We discuss the new but traditional osteosynthesis method.A surgical suture osteosynthesis could act as an additional alternative when special fractures or conditions occur. Postoperative results in early period are satisfactory. Long-term results and clinical experience should be analyzed and discussed.

To investigate the antimicrobial resistance among the nosocomial gram-negative pathogens from 15 teaching hospitals located in different areas in China in 2005.A total of 1927 non-repetitive nosocomial gram-negative pathogens were collected from 15 teaching hospitals in different areas in China and sent to the central lab for reidentification and susceptibility testing. The levels of minimal inhibitory concentration (MIC) of 18 antimicrobial agents were determined by agar dilution method. WHONET 5.4 software was used to analyze the data.The strains of Escherichia coli, Klebsiella pneumoniae, and Proteous mirabilis isolates that did not produce extended spectrum beta lactamases (ESBLs) showed high sensitivity to beta-lactams. The antibiotics with a susceptibility rates over 80% against the strains of Entorobacter cloacae, Enterobacter aerogene, Citrobacter spp, Serratia spp, and Proteous vulgaris producing AmpC enzyme included meropenem, imipenem, and piperacillin-tazobactam, and these 3 drugs showed a susceptibility rate of more than 80% against the ESBL-producing strains of Escherichia coli and Klebsiella pneumoniae. Other antimicrobial agents showing a relatively high activity against Enterobacter spp, Citrobacter spp, Serratia spp and Proteous vulgaris included cefepime (67.3% - 100%), amikacin (67.3% - 95.2%), ceftazidime (52.9% - 100%) and cefoperazone-sulbactam (51.9% - 100%). The susceptibility rate of fluoroquinolones was 34.8% - 36.1% against non-ESBL-producing Escherichia coli and was 13.4% - 17.1% against ESBL-producing isolates. The most active agent against Pseudomonas aeruginosa was polymyxin B (95.6%). The agents with the activity rates of 70% - 80% included meropenem, imipenem, amikacin, and piperacillin-tazobactam. The antibiotic with a high susceptible rate against Acinetobacter baumannii was polymyxin B (98.3%), followed by imipenem (80.8%), meropenem (76.2%), and minocycline (67.4%). The susceptible rates of other agents were all below 60%. The agents with relatively high activity against Stenotrophomonas maltophilia included minocycline (85%), levofloxacin (82.5%), and trimethoprim-sulfamethoxazole (77.5%). The agents with a relatively high activity against Burkholderia cepacia included minocycline (77.2%) and meropenem (61.4%).Carbapenem, piperacillin-tazobactam, amikacin and cefepime remained relatively high activity against nosocomial Enterobacteriaceae, Non-fermenting pathogens have lower susceptibility to the antimicrobial agents than before.

Double antibody sandwich-type ELISA was used to detect rhG-CSF in serum to study the pharmacokinetics of rhG-CSF, PEG-rhG-CSF and rHSA-hG-CSF in mice and to confirm that PEGlyation and albumin fusion of rhG-CSF technology can prolong half-life of G-CSF. Pharmacokinetic parameters were calculated with 3P87 software. T1/2 s of rhG-CSF, PEG-rhG-CSF and rHSA-hG-CSF are 2. 1 , 14.2 and 10. 6 h, respectively. T1/2 s of PEG- rhG-CSF and rHSA-hG-CSF are 7, 5 times than T1/2 s of rhG-CSF, respectively. Tpeak s of PEG-rhG-CSF and rHSA-hG-CSF are 15, 13 times than Tpeak of rhG-CSF, respectively. The result of ELISA indicates that PEGlyation and albumin fusion of rhG-CSF technology can prolong half-life of G-CSF.

To search for a more ideal way to disinfect infectious root canal of deciduous tooth.According to bacterial occurrence and clinical effect, 40 cases of infectious root canal of deciduous teeth disinfected with Ca(OH)2 were investigated, the other 40 cases with traditional way of using FC and CP respectively, were used as the control.Fist of all, the bacteria of deciduous tooth infectious root canal were same as the bacteria of permanent tooth infectious root canal. Secondly, Ca(OH)2 was stronger in disinfecting deciduous tooth infectious root canal. Thirdly, so far as sterilization was concerned, Ca(OH)2 had the same disinfectant as FC, and was better than CP, but FC had some irritation and poisonous side-effects to periodical tissue. So Ca(OH)2 was safe and effective for deciduous tooth infectious root canal.It was a more ideal way to disinfect deciduous tooth infectious root canal by Ca(OH)2.

Osteonecrosis of the femoral head (ONFH) can be caused by a decrease in the activity or numbers of osteoblasts, a process in which apoptosis may play an essential role. We investigated the effect of dexamethasone (Dex) combined with hypoxic stress on murine osteoblastic MC3T3-E1 cells. Flow cytometry, western blot and real-time quantitative PCR analyses revealed that hypoxia significantly enhanced Dex-induced apoptosis. Further data demonstrated that both the death receptor and the mitochondria-mediated pathway were involved in Dex-induced apoptosis under hypoxic conditions. However, the death receptor pathway had only a minor effect on this process. The expression levels of Bcl-2 and Bax, which regulate the mitochondria-initiated apoptotic cascade signaling pathway, were significantly different in response to Dex and hypoxia. The mitochondrial membrane potential collapsed, and the inhibitor brain- derived neurotrophic factor (BDNF) conferred effective protection against apoptosis. In summary, the mitochondria-mediated apoptotic pathway functions in osteoblast apoptosis that is induced by Dex in a hypoxic environment, and the present study may help us to gain further insight into the molecular mechanisms of steroid-induced ONFH.